Compositions and methods for treating and preventing proteinuria and endothelial erosion

ABSTRACT

In some embodiments, a system and/or method may inhibit and/or ameliorate proteinuria and related comorbidities in a subject. In some embodiments, proteinuria and/or Endothelial Erosion (EE)™ and related comorbidities may be inhibited and/or ameliorated by adjusting a subject&#39;s zeta potential to at least within acceptable levels associated with a healthy subject. A subject&#39;s zeta potential may be adjusted by administering pharmaceutical compositions which: increases a negative electrostatic potential of various tissues in a subject; increases the negative electrical surface charges of a subject&#39;s blood cells, plasma proteins and/or glycocalyx; and/or adjusts various properties of a subject&#39;s bloodstream (e.g., the blood serum pH). In some embodiments, Proteinuria and/or Endothelial Erosion (EE)™ and related comorbidities may be inhibited and/or ameliorated by: adjusting a subject&#39;s blood serum pH; repairing damage to the glycocalyx in a subject; and/or increasing blood flow, supply essential elements and/or replenish essential elements lost through proteinuria.

CROSS REFERENCE TO RELATED APPLICATIONS

This application is a divisional of Ser. No. 16/173,475, filed Oct. 29,2018, which claims priority to U.S. Provisional Patent Application No.62/578,076 filed on Oct. 27, 2017, both entitled “COMPOSITIONS ANDMETHODS FOR TREATING AND PREVENTING PROTEINURIA AND ENDOTHELIALEROSION”. The Provisional Application and the Copending Non-provisionalApplication are hereby incorporated by reference in their entireties.

BACKGROUND OF THE INVENTION Field of the Invention

The present disclosure generally relates to proteinuria, EndothelialErosion (EE)™ and related metabolic, vascular and neurologicalcomorbidities. More particularly, the disclosure generally relates tocompositions and methods for treating and preventing proteinuria,Endothelial Erosion (EE)™ and related metabolic, vascular andneurological comorbidities.

Description of the Relevant Art

The glomerular capillary wall of a healthy kidney functions as a barrierto prevent plasma proteins from entering the urine, based on the sizeand electrical surface charge of the plasma proteins. The primarybarrier for ultrafiltration of plasma in renal glomeruli comprises fourlayers (e.g., the glycocalyx, a fenestrated endothelium, a glomerularbasement membrane, and diaphragms located between the foot processes ofthe podocytes). If the glomerulus is intact and functioning properly,only trace amounts of albumin and other large plasma proteins escapeinto the glomerular filtrate. Defective glomerular filtration is acommon condition associated with a large number of acquired andinherited diseases. Defective glomerular filtration may result in theleakage, to varying degrees, of plasma albumin and other large plasmaproteins, leading to protein in the urine (proteinuria), and aninsufficient quantity of essential plasma proteins in the bloodstream(hypoproteinemia), and progressive, degenerative kidney disease withmany related metabolic, vascular and neurological comorbidities. MinimalChange Disease (MCD) is an example of a disease characterized by suddenand massive proteinuria and hypoproteinemia, and a rapid progression toEnd Stage Renal Disease (ESRD) with many related metabolic, vascular andneurological comorbidities (peripheral artery disease, peripheralneuropathy, retinopathy, systemic scarring, sclerosis, fibrosis, etc.).

Kidney disease is the ninth leading cause of death in the United States.It is estimated that over 31 million Americans (10% of the adultpopulation) suffer from chronic kidney disease (CKD). However, mostpeople with chronic kidney disease will not reach end stage renaldisease (ESRD) because cardiac failure and/or some other cardiovascularrelated comorbidity (e.g. cardio-renal syndrome or renal-cardiosyndrome) will kill them before the progression to ESRD can take place.There is a well-established clinical relationship between pathologicalstates of the kidney and of the heart which displays the link betweenchronic kidney disease and cardiovascular disease; i.e., the severity ofproteinuria can predict the rate of progression of cardiovasculardisease as well as chronic kidney disease. One important determinant ina given person's rate of progression of chronic kidney disease and rateof progression of cardiovascular disease and related comorbidities isthe amount of protein in the urine (“proteinuria”) and the amount ofprotein in the serum (low albumin levels are referred to as“hypoalbuminemia,” and hypoalbuminemia can be a better predictor ofmortality rates in “otherwise healthy” subjects than even severeobesity). Generally, patients with greater amounts of proteinuria have amore aggressive form of chronic kidney disease as well as a moreaggressive form of cardiovascular disease and related comorbidities tendto present with increased severity as well. For this reason, manyresearchers have attempted to design treatments to reduce and preventthe excess amounts of protein in urine. Amongst patients who haveproteinuria, there is a broad range of excess protein that may beexcreted. This range of excess protein loss through the urine caninclude small losses in conditions such as microalbuminuria (which ischaracterized by the excretion of 30-300mg albumin in the urine during a24-hour period) as well as larger losses, as seen in nephrotic syndrome(which is characterized by the excretion of over 3.5 g of protein in theurine during a 24-hour period) and any amount of protein excreted in theurine which falls in between these two conditions.

Accordingly, there exists a need, heretofore unfulfilled, for treatmentmethods that inhibit and/or ameliorate proteinuria.

SUMMARY

In some embodiments, a system and/or method may inhibit and/orameliorate proteinuria and related comorbidities in a subject. In someembodiments, a system and/or method may inhibit and/or ameliorateEndothelial Erosion (EE) and related comorbidities in a subject.Proteinuria and/or Endothelial Erosion (EE) and related comorbiditiesmay be inhibited and/or ameliorated by: maximizing a subject's serumzeta potential; repairing damage to the endothelial surface layer in asubject; making up for a subject's urinary losses of plasma proteins;and/or optimizing blood flow and facilitating healing of damage causedby Proteinuria and/or Endothelial Erosion (EE). A subject's serum zetapotential may be maximized by administering pharmaceutical compositionswhich: optimize the negative electrical surface charges of a subject'sblood cells, plasma proteins, and/or endothelial surface layer; optimizethe composition of a subject's bloodstream (e.g., ionic strength and ionconcentrations), and/or optimize the properties of the subject'sbloodstream (e.g., serum pH). In some embodiments, proteinuria and/orEndothelial Erosion (EE) and related comorbidities may be inhibitedand/or ameliorated by administering pharmaceutical compositions whichinhibit and/or ameliorate (e.g., maintain, repair and/or regrow theglycocalyx) damage to the endothelial surface layer in a subject.Proteinuria and/or Endothelial Erosion (EE) and related comorbiditiesmay be inhibited and/or ameliorated by administering pharmaceuticalcompositions which increase blood flow, supply essential elements and/orreplenish essential elements lost through proteinuria.

In some embodiments, a method may include treating proteinuria. Themethod may include adjusting a subject's serum zeta potential. Themethod may include increasing the health of a subject's endothelialsurface layer. The method may include replenishing a subject's urinarylosses of plasma proteins. The method may include ameliorating damage toa subject's glomerulus.

In some embodiments, the subject's serum zeta potential is a potentialfor a subject's blood cells, plasma proteins, and endothelial surfacelayer to electrostatically repel one another.

In some embodiments, adjusting a subject's serum zeta potential includesincreasing the subject's serum zeta potential. Adjusting a subject'sserum zeta potential may include maximizing the subject's serum zetapotential. Adjusting a subject's serum zeta potential may includeelectrostatically inhibiting coagulation in the subject's blood serum.Adjusting a subject's serum zeta potential may include administering acomposition to the subject comprising sodium chloride, potassiumchloride, magnesium chloride, potassium citrate, and/or olive leafextract. Adjusting a subject's serum zeta potential may includeadjusting a pH of the subject's serum. Adjusting a subject's serum zetapotential may include adjusting a pH of the subject's serum to betweenabout 7.35 and about 7.45.

In some embodiments, increasing the health of subject's endothelialsurface layer may include ameliorating damage to the endothelial surfacelayer. Increasing a health of subject's endothelial surface layer mayinclude administering a composition to the subject which includescompounds used by the subject's own endothelial cells to form thesubject's endothelial glycocalyx. Increasing the health of a subject'sendothelial surface layer may include administering a composition to thesubject which includes hyaluronic acid, N-acetyl glucosamine,glucosamine sulfate, chondroitin sulfate, n-acetyl cysteine, and/ormethylsulfonylmethane.

In some embodiments, replenishing urinary losses of plasma proteins ofthe subject may include administering a composition to the subject whichincludes lactoferrin and sources of sodium, chloride, potassium, andmagnesium.

In some embodiments, ameliorating damage to a subject's glomerulus mayinclude administering a composition to the subject which includestocotrienols, nattokinase, n-acetyl cysteine, and methylsulfonylmethane.Tocotrienols may include delta and gamma tocotrienols. Tocotrienols mayinclude delta and gamma tocotrienols in a 90:10 ratio respectively.Damage to a subject's glomerulus may include scarring and/or sclerosis.

In some embodiments, a chemical composition may be configured to treatproteinuria. The composition may include a first group of compoundsproviding bioavailable sources of sodium, potassium, chloride, oliveleaf extract, potassium citrate, and/or magnesium. The first group ofcompounds may adjust a subject's serum zeta potential. The compositionmay include a second group of compounds including hyaluronic acid,n-acetyl glucosamine, n-acetyl cysteine, methylsulfonylmethane,glucosamine sulfate and/or chondroitin sulfate. The second group ofcompounds may increase the health of a subject's endothelial surfacelayer. The composition may include a third group of compounds includingbioavailable sources of magnesium, sodium, potassium, chloride, and/orlactoferrin. The third group of compounds may replenish a subject'surinary losses of plasma proteins. The composition may include a fourthgroup of compounds comprising nattokinase, methylsulfonylmethane,n-acetyl cysteine and/or tocotrienols. The fourth group of compounds mayameliorate damage to a subject's glomerulus.

In some embodiments, the first group of compounds provides bioavailablesources of sodium, potassium, chloride, olive leaf extract, potassiumcitrate, and magnesium.

In some embodiments, the second group of compounds includes hyaluronicacid, n-acetyl glucosamine, n-acetyl cysteine, methylsulfonylmethane,glucosamine sulfate and chondroitin sulfate.

In some embodiments, the third group of compounds may includebioavailable sources of magnesium, sodium, potassium, chloride, andlactoferrin.

In some embodiments, the fourth group may include nattokinase,methylsulfonylmethane, n-acetyl cysteine, and tocotrienols.

BRIEF DESCRIPTION OF THE DRAWINGS

Advantages of the present invention may become apparent to those skilledin the art with the benefit of the following detailed description of thepreferred embodiments and upon reference to the accompanying drawings.

FIGS. 1A, 1B, 1C, and 1D depict a pictorial representation of zetapotential and the particles' ability in solution to electrostaticallyrepel one another.

FIG. 2 depicts an embodiment of a portion of a subject's healthy andproperly functioning cardiovascular system coupled to a healthy kidneywith a healthy Zeta Shield (a healthy glycocalyx and a healthy zetapotential) with no Proteinuria and no Endothelial Erosion.

FIGS. 3 and 4 depict an embodiment of a portion of a subject's unhealthycardiovascular system shortly after experiencing a sudden, massive lossof zeta potential as typically seen in Minimal Change Disease (MCD) andPre-eclampsia. The loss of zeta potential causes Proteinuria,hypercoagulation and hypertension. The Proteinuria causeshypoproteinemia which leads to progressive systemic ischemia, edema,oxidative stress and iron deficiency anemia. At this initial stage, thesubject's glycocalyx is still intact and there is no EndothelialErosion.

FIGS. 5, 6, 7, 8, 9, 10, 11, 12, 13, 14 and 15 depict an embodiment of aportion of a subject's unhealthy cardiovascular system showing theprogression of Endothelial Erosion caused by hydraulic friction due tothe loss of the subject's Zeta Potential. Endothelial Erosion beginswith the erosion of the glycocalyx (FIG. 4) and ends in catastrophicfailure or thrombosis (FIG. 15) causing a heart attack, stroke orpulmonary embolism.

FIG. 16 depicts a pictorial summary of a theory of chronic degenerativedisease over an average human lifespan.

FIG. 17 depicts an embodiment of a cross-section of a subject'sunhealthy blood vessel damaged by hydraulic friction and EndothelialErosion due to the loss of the subject's Zeta Potential.

FIG. 18 depicts an embodiment of a cross-section of a subject's healthyblood vessel protected from hydraulic friction and Endothelial Erosionby a healthy Zeta Shield (a healthy glycocalyx and a healthy zetapotential strong enough to electrostatically repel blood cells andplasma proteins).

FIG. 19 depicts a graph of the average human lifespan for the at leastthe last several hundred years and how it has improved and could improveeven more.

FIGS. 20-21 depict a spread sheet showing the preliminary resultsachieved by treating a cat with severe proteinuria with compositionsdescribed herein.

While the invention is susceptible to various modifications andalternative forms, specific embodiments thereof are shown by way ofexample in the drawings and may herein be described in detail. Thedrawings may not be to scale. It should be understood, however, that thedrawings and detailed description thereto are not intended to limit theinvention to the particular form disclosed, but on the contrary, theintention is to cover all modifications, equivalents and alternativesfalling within the spirit and scope of the present invention as definedby the appended claims.

The headings used herein are for organizational purposes only and arenot meant to be used to limit the scope of the description. As usedthroughout this application, the word “may” is used in a permissivesense (i.e., meaning having the potential to), rather than the mandatorysense (i.e., meaning must). The words “include,” “including,” and“includes” indicate open-ended relationships and therefore meanincluding, but not limited to. Similarly, the words “have,” “having,”and “has” also indicated open-ended relationships, and thus mean having,but not limited to. The terms “first,” “second,” “third,” and so forthas used herein are used as labels for nouns that they precede, and donot imply any type of ordering (e.g., spatial, temporal, logical, etc.)unless such an ordering is otherwise explicitly indicated. For example,a “third chemical compound added to a pharmaceutical composition” doesnot preclude scenarios in which a “fourth chemical compound added to apharmaceutical composition” is added prior to the third chemicalcompound, unless otherwise specified. Similarly, a “second” feature doesnot require that a “first” feature be implemented prior to the “second”feature, unless otherwise specified.

Various components may be described as “configured to” perform a task ortasks. In such contexts, “configured to” is a broad recitation generallymeaning “having structure that” performs the task or tasks duringoperation. As such, the component can be configured to perform the taskeven when the component is not currently performing that task. In somecontexts, “configured to” may be a broad recitation of structuregenerally meaning “having chemical compounds that” perform the task ortasks during use. As such, the component can be configured to performthe task even when the component is not currently being used.

Various components may be described as performing a task or tasks, forconvenience in the description. Such descriptions should be interpretedas including the phrase “configured to.” Reciting a component that isconfigured to perform one or more tasks is expressly intended not toinvoke 35 U.S.C. § 112, paragraph six, interpretation for thatcomponent.

The scope of the present disclosure includes any feature or combinationof features disclosed herein (either explicitly or implicitly), or anygeneralization thereof, whether or not it mitigates any or all of theproblems addressed herein. Accordingly, new claims may be formulatedduring prosecution of this application (or an application claimingpriority thereto) to any such combination of features. In particular,with reference to the appended claims, features from dependent claimsmay be combined with those of the independent claims and features fromrespective independent claims may be combined in any appropriate mannerand not merely in the specific combinations enumerated in the appendedclaims.

It is to be understood the present invention is not limited toparticular devices or biological systems, which may, of course, vary. Itis also to be understood that the terminology used herein is for thepurpose of describing particular embodiments only, and is not intendedto be limiting. As used in this specification and the appended claims,the singular forms “a”, “an”, and “the” include singular and pluralreferents unless the content clearly dictates otherwise. Thus, forexample, reference to “a linker” includes one or more linkers.

DETAILED DESCRIPTION Definitions

Unless defined otherwise, all technical and scientific terms used hereinhave the same meaning as commonly understood by one of ordinary skill inthe art.

The terms “administration,” “administering,” or the like, as used hereinwhen used in the context of providing a pharmaceutical, cosmeceutical ornutraceutical composition to a subject generally refers to providing tothe subject one or more pharmaceutical, “over-the-counter” (OTC) ornutraceutical compositions in combination with an appropriate deliveryvehicle by any means such that the administered compound achieves one ormore of the intended biological effects for which the compound wasadministered. By way of non-limiting example, a composition may beadministered parenteral, subcutaneous, intravenous, intracoronary,rectal, intramuscular, intra-peritoneal, transdermal, or buccal routesof delivery. Alternatively, or concurrently, administration may be bythe oral route. The dosage of pharmacologically active compound that isadministered will be dependent upon multiple factors, such as the age,health, weight, and/or disease state of the recipient, concurrenttreatments, if any, the frequency of treatment, and/or the nature andmagnitude of the biological effect that is desired.

The term “ameliorating,” as used herein, when used in the context ofmodulating a pathological or disease state, generally refers to reducingor treating damage resulting from the disease state. In some contexts,the term may generally refer to addressing in a positive manner one ormore root causes of a disease state.

The term “animal” as used herein generally refers to any member of thekingdom Animalia, comprising multicellular organisms that have awell-defined shape and usually limited growth, can move voluntarily,actively acquire food and digest it internally, and have sensory andnervous systems that allow them to respond rapidly to stimuli: someclassification schemes also include protozoa and certain othersingle-celled eukaryotes that have motility and animal like nutritionalmodes. Generally the term animal as used herein does not refer tohumans.

The term “anti-inflammatory” as used herein generally refers to asubstance acting to reduce certain signs of inflammation (e.g.,swelling, tenderness, fever, and pain).

The term “connected” as used herein generally refers to pieces which maybe joined or linked together.

The term “coupled” as used herein generally refers to pieces which maybe used operatively with each other, or joined or linked together, withor without one or more intervening members.

The term “directly” as used herein generally refers to one structure inphysical contact with another structure, or, when used in reference to aprocedure, means that one process effects another process or structurewithout the involvement of an intermediate step or component.

The terms “effective concentration” or “effective amount” as used hereingenerally refers to a sufficient amount of the pharmaceutically activeagent that is added to decrease, prevent or inhibit or treat one ormaladies described herein and/or related conditions. The amount willvary for each compound and upon known factors related to the item or useto which the pharmaceutically active agent is applied.

The terms “in need of treatment” or “in need thereof” when used in thecontext of a subject being administered a pharmacologically activecomposition, generally refers to a judgment made by an appropriatehealthcare provider that an individual or animal requires or willbenefit from a specified treatment or medical intervention. Suchjudgments may be made based on a variety of factors that are in therealm of expertise of healthcare providers, but include knowledge thatthe individual or animal is ill, will be ill, or is at risk of becomingill, as the result of a condition that may be ameliorated or treatedwith the specified medical intervention.

The term “malady” as used herein generally refers to any disorder ordisease of the body or any undesirable or disordered conditionincluding, but not limited to, illness, sickness, affliction, complaint,ailment, indisposition, virus, disease, fungus, infection, disease, etc.

Terms such as “pharmaceutical composition,” “pharmaceuticalformulation,” “pharmaceutical preparation,” or the like, are used hereinto generally refer to formulations that are adapted to deliver aprescribed dosage of one or more pharmacologically active compounds to acell, a group of cells, an organ or tissue, an animal or a human.Methods of incorporating pharmacologically active compounds intopharmaceutical preparations are widely known in the art. Thedetermination of an appropriate prescribed dosage of a pharmacologicallyactive compound to include in a pharmaceutical composition in order toachieve a desired biological outcome is within the skill level of anordinary practitioner of the art. A pharmaceutical composition may beprovided as sustained-release or timed-release formulations. Suchformulations may release a bolus of a compound from the formulation at adesired time, or may ensure a relatively constant amount of the compoundpresent in the dosage is released over a given period of time. Termssuch as “sustained release,” “controlled release,” or “timed release”and the like are widely used in the pharmaceutical arts and are readilyunderstood by a practitioner of ordinary skill in the art.Pharmaceutical preparations may be prepared as solids, semi-solids,gels, hydrogels, liquids, solutions, suspensions, emulsions, aerosols,powders, or combinations thereof. Included in a pharmaceuticalpreparation may be one or more carriers, preservatives, flavorings,excipients, coatings, stabilizers, binders, solvents and/or auxiliariesthat are, typically, pharmacologically inert. It will be readilyappreciated by an ordinary practitioner of the art that, included withinthe meaning of the term are pharmaceutically acceptable salts ofcompounds. It will further be appreciated by an ordinary practitioner ofthe art that the term also encompasses those pharmaceutical compositionsthat contain an admixture of two or more pharmacologically activecompounds, such compounds being administered, for example, as acombination therapy.

A “pharmaceutically or nutraceutically acceptable formulation,” as usedherein, generally refers to a non-toxic formulation containing apredetermined dosage of a pharmaceutical and/or nutraceuticalcomposition, wherein the dosage of the pharmaceutical and/ornutraceutical composition is adequate to achieve a desired biologicaloutcome. The meaning of the term may generally include an appropriatedelivery vehicle that is suitable for properly delivering thepharmaceutical composition in order to achieve the desired biologicaloutcome.

The term “pharmacologically inert,” as used herein, generally refers toa compound, additive, binder, vehicle, and the like, that issubstantially free of any pharmacologic or “drug-like” activity.

The terms “reducing” and “inhibiting,” as used herein, when used in thecontext of modulating a pathological or disease state, generally refersto the prevention and/or reduction of at least a portion of the negativeconsequences of the disease state. When used in the context of anadverse side effect associated with the administration of a drug to asubject, the term(s) generally refer to a net reduction in the severityor seriousness of said adverse side effects.

The term “subject” as used herein generally refers to a mammal (e.g.,felines, canines), and in particular to a human.

The term “colloid” as used herein generally refers to a mixture in whichone substance of dispersed insoluble particles is suspended throughout asolution (e.g., blood is a colloid).

The term “plasma” as used herein generally refers to a constituent ofthe blood of an animal or a subject which can be separated from the redblood cells and clotting factors present in the blood stream of a livinganimal or subject.

The term “serum” as used herein generally refers to the entirety offlowing blood within the vasculature of a living animal or subject.

The phrase “therapeutically effective amount” generally refers to anamount of a drug or pharmaceutical composition that will elicit at leastone desired biological or physiological response of a cell, a tissue, asystem, animal or human that is being sought by a researcher,veterinarian, physician or other caregiver.

The phrase “zeta potential” generally refers to a measurement of theability or the potential of insoluble particles in a colloidal solutionto electrostatically repel one another. This may be dependent on severalfactors including the negative electrical surface charge of theparticles and various properties of the solution (e.g., the pH). Thezeta potential may generally refer to the potential difference existingbetween the surface of insoluble particles in a colloidal solution. Zetapotential was initially discovered in 1879 by German scientist Hermannvon Helmholtz, and was influential in Einstein's 1905 paper aboutBrownian motion. A direct measurement of the serum zeta potential of aliving organism is not feasible at the present time because themeasurement device will collapse the fluid during the measurementprocess, but this difficulty with direct measurement does not make itimpossible to indirectly measure a subject's or an animal's serum zetapotential. Researchers commonly use zeta potential calculations todesign self-emulsifying drug delivery systems (SEDDS) for use in animalsand/or subjects. In the same way, researchers are able to use indirectmeasurements of serum zeta potential to quantify results and identifychemical steps to maximize, improve, or optimize the serum zetapotential of an animal or subject.

The term “serum zeta potential” refers to the ability or the potentialfor a subject's blood cells and plasma proteins to electrostaticallyrepel one another, and the ability or the potential for the subject'sendothelial surface layer to electrostatically repel the subject's bloodcells and plasma proteins. A subject's serum zeta potential may bedependent on several factors including the negative electrical surfacecharges of the subject's endothelial surface layer, blood cells, andplasma proteins, as well as various properties of the subject'sbloodstream (e.g., the blood serum pH). The serum zeta potential can beaffected, modified, collapsed or enhanced by a large number of widelydivergent phenomena, most notably including: serum volume, serumcomposition, environmental factors (e.g., infectious disease, thepresence of toxins, the physiological state of the subject or animalwhich may be active or at rest, etc.).

The term “Van der Waals Forces” refers to the tendency of insolubleparticles in a solution to clump together and settle at the bottom ofthe container. In situations where a subject or animal's serum zetapotential suddenly collapses, environmental factors are said to haveenabled the Van der Waals Forces to overcome the serum zeta potential,resulting in a loss of the ability of the negatively electricallycharged components (e.g., plasma proteins, blood cells, and/orendothelial surface layer, etc.) of the subject or animal's serum torepel one another.

The phrase “maximize serum zeta potential” refers to a process in whicha subject or animal is treated with compositions of a pharmaceuticaland/or nutraceutical nature. These compositions are designed to impactthe viscosity, volume, composition, and negative electrical surfacecharge of the components of the serum of a subject or animal such thatthe Van der Waals Forces between these components of the serum areovercome by the zeta potential of the serum of a subject or animal.Proteinuria is currently the best available proximate biomarker by whichto ascertain the level of a subject or animal's serum zeta potentialbecause the normal function of the glomerulus is not possible when serumzeta potential is reduced. Events such as proteinuria after exercise orproteinuria in conjunction with hyperglycemia are direct evidence thatthe characteristics of the serum are as critical to the normal functionof the glomerulus as the structural integrity of the glomerulus itself;hence to maximize the serum zeta potential is to enhance the aspects ofthe serum which allow for normal renal function and circulation ofserum.

The phrase “endothelial surface layer (ESL)” refers to the region of thevasculature which enables selective endothelial boundary permeability.There are three main parts of the endothelial surface layer, and each ofthem undergoes consistent modification in normal, ordinary life for bothanimals and subjects. The three parts of the ESL consist of: theendothelial glycocalyx, which is a hair-like growth produced by theendothelial cells comprised of heparan sulfate, hyaluronic acid,proteoglycans and glycosarninoglycans; objects and particles whichbecome attached to the endothelial glycocalyx such as albumin and metalsin circulation; and the boundary region, which is produced by thenegative electric surface charge of the endothelial glycocalyx and whichprovides the negative electric potential needed to prevent erythrocytesand other large, insoluble serum particles from directly coming intocontact with the endothelial glycocalyx, which would result in erosion.Albumin is a common protein in the human serum (typical concentrationsrange from 3.5-5.0 g/dL) and it is commonly found attached to theendothelial glycocalyx, as this is a normal part of metabolism for asubject and/or an animal in which the albumin molecule is transcytosedthrough the endothelial boundary into the interstitial space. The ESL istherefore a dynamic part of the physiology of a subject or an animalwhich needs continuous maintenance at the cellular level to ensure thehealth of a subject or an animal.

The term “Zeta Shield” is intended as a trademark and generally refersto an electromagnetic shield that lines all healthy blood vessels andprotects the subject's endothelial cells and their associatedendothelial surface layer from hydraulic friction and EndothelialErosion (EE). The strength of a subject's Zeta Shield is dependent onthe subject's serum zeta potential as well as the health of theendothelial surface layer.

The term “Endothelial Erosion (EE)” is intended as a trademark and togenerally refer herein to hydraulic friction which damages or wears awayand erodes the endothelial barrier (which consists of the basementmembrane, the endothelial cells, and the endothelial surface layerincluding the endothelial glycocalyx) and inhibits the properfunctioning of the vascular endothelium.

Embodiments

In some embodiments, systems presented herein may allow for inhibitionand/or amelioration of proteinuria. It has been accepted by many in themedical field that the kidneys are basically size selective filters andthat proteinuria is a result of inflammation and/or a result of anauto-immune disorder. Proteinuria is thought to be a result ofinflammation causing kidney tissue to become inflamed and expand causingopenings to expand and allowing plasma proteins to pass through.

However, endothelial cell fenestrations are larger than plasma proteinsand as such if the kidney was a simple mechanical filter then plasmaproteins would flow through even healthy kidneys. The mechanical filtermodel for kidneys does not explain sudden onset proteinuria, suddenfailure of kidney transplants, the elevated sedimentation rate orelevated blood pressure associated with proteinuria, or minimal changenephrotic syndrome.

What does explain the inconsistencies of a mechanical filter model forkidneys is that the kidney is actually an electrostatic filter such thatproteinuria is actually an electromagnetic problem. Everything desirablein your bloodstream has a negative electrical surface charge (or istransported by elements with a negative electrical surface charge suchas red blood cells or plasma proteins) while the wastes typicallyfiltered out by the kidneys do not have a negative electrical surfacecharge. While the kidney membranes have a negative electrical surfacecharge and as such electrostatically repel, for example, plasma proteinsand red blood cells. Unwanted waste is typically uncharged and as suchpasses through endothelial fenestrations and expelled through theurinary tract. Proteinuria therefore occurs at least in part when kidneymembranes lose some of their electrostatic surface charge and hencetheir ability or potential to electrostatically repel plasma proteinsallowing the proteins to pass through the kidneys and into the urinarytract.

Further evidence can be seen for the electrokinetic model of kidneyfunction when one looks at hypercoagulation in a subject's blood(erythrocyte sedimentation rate is a common test for hypercoagulation)that sometimes accompanies proteinuria. This hypercoagulation is thedirect result of the loss of a subject's serum zeta potential, or theability/ potential for the subject's red blood cells and plasma proteinsto electrostatically repel one another. FIGS. IA-B depicts a pictorialrepresentation of serum zeta potential and the particles' ability insolution to electrostatically repel one another. As is depicted in FIGS.IA-B the particles 50 have an appropriate charge which allows theparticles to repel one another in solution and therefore remaindispersed and resist coagulation. As is depicted in FIGS. IC-D theparticles 50 do not have an appropriate charge which allows theparticles to repel one another in solution and therefore the particlesbegin to clump together and coagulate 60.

When there is a high zeta potential in a colloidal solution, particlesdistribute evenly over time throughout the solution due to the particleselectrostatically repelling each other. When there is a low zetapotential in a colloidal solution, particles will clump together andsettle to the bottom of the solution due to the particles no longerelectrostatically repelling each other. Zeta potential may be dependenton the negative electrical surface charge of the particles as well asvarious properties of the solution itself (e.g., pH). The endothelialbarrier has a negative electrical surface charge which allows for evenfluid flow of the blood throughout the circulatory system by repulsingnegatively charged blood cells and plasma proteins (e.g., reducinghydraulic friction, reducing blood pressure and reducing wear againstthe endothelial barrier such that subject's with a high zeta potentialeffectively have blood flowing with little or no friction).

The vascular endothelium lining the inner surface of blood vesselsserves as an interface for circulating blood components to interact withcells of the vascular wall and surrounding extravascular tissues. Afunction of vascular endothelia, especially those in exchangemicrovessels (capillaries and postcapillary venules), is to provide asemipermeable barrier that controls blood-tissue exchange of fluids,nutrients, and metabolic wastes while at the same time preventingpathogens or harmful materials in circulation from entering intotissues. Plasma leakage due to disruption and/or damage of vascularendothelia disturbs fluid homeostasis and impairs tissue oxygenation, apathophysiological process contributing to multiple organ dysfunctionassociated with trauma, infection, metabolic disorder, and other formsof disease. FIG. 2 depicts an embodiment of a portion of a healthysubject's cardiovascular system coupled to a healthy kidney with ahealthy Zeta Shield (a healthy glycocalyx and a healthy zeta potential)and how a subject's systems are supposed to function when healthy.Healthy red blood cells and plasma proteins (70) have a negativeelectrical surface charge which allows them to electrostatically repelone another and disperse evenly throughout an organism's bloodstream. Ahealthy endothelial barrier (80) also has a negative electrical chargewhich allows it to electrostatically repel red blood cells and plasmaproteins; this process thus reduces hydraulic friction, blood pressureand wear against the endothelial surface layer. A healthy glomerularfiltration barrier (GFB) (90) also has a negative electrical charge soit electrostatically repels red blood cells and plasma proteins, therebyelectrostatically separating the waste in a subject's bloodstream fromthe negatively charged red blood cells and plasma proteins.

Similarly, as a subject's Zeta potential is reduced several negativeeffects result. Red blood cells begin to clump together and are nolonger electrostatically repelled by the endothelial barrier resultingin increasing hydraulic friction, blood pressure, and wear against theendothelial barrier (“Endothelial Erosion™”). Endothelial Erosion™results from constant hydraulic friction and wear against theendothelial barrier beginning with erosion of the Glycocalyx.Endothelial erosion can eventually lead to endothelial barrier failureand breach. In the same manner, the kidney filter membranes lose theirability to electrostatically repel plasma proteins resulting in theproteins passing through the kidney filter membranes along with wasteproducts (“Proteinuria”).

FIGS. 3-4 depict an embodiment of a portion of an unhealthy subject'scardiovascular system shortly after experiencing a sudden, massive lossof serum zeta potential as typically seen in Minimal Change Disease(MCD) and Pre-eclampsia. The loss of serum zeta potential causesProteinuria I 00, hypercoagulation and hypertension. Proteinuria leadsto hypoproteinemia 110, a condition in which an insufficient quantity ofplasma proteins remain to transport nutrients to cells or to pull thewaste products from cells into the subject's bloodstream. If thehypoproteinemia continues unabated, cells throughout the subject will bestressed and can begin to die from toxicity and nutrient deficiencycommonly referred to as ischemia 130. As hypoproteinemia continues,cellular waste continues to collect in the subject's tissues causingsevere swelling or edema 120. Due to urinary losses of transferrin andceruloplasmin, oxidative stress 140 and iron deficiency anemia 140 canresult. One or more of these conditions may lead to the liver respondingby over-producing cholesterol, a condition commonly referred to ashyperlipidemia.

FIGS. 5-15 depict an embodiment of a portion of an unhealthy subject'scardiovascular system with a depleted zeta potential and a secondpathway leading to catastrophic failure of the subject's body's systemsresulting from loss of the subject's serum zeta potential over anextended period of time. The second pathway typically extends over aperiod of decades and currently is thought to be normal conditions orailments associated with aging. The second pathway may begin with redblood cells coagulating due to a loss of serum zeta potential. Thecoagulated blood cells rub against the endothelial barrier, increasinghydraulic friction, blood pressure, and wear of the endothelial barrier.The process begins with glycocalyx erosion 210 from the frictiongenerated by clumps of coagulated blood cells penetrating into theendothelial surface layer, causing damage to the endothelial glycocalyxand damaging the endothelial cells (e.g., see FIG. 5). Glycocalyxerosion leads to Endothelial Erosion (EE), which includes endothelialcell damage and endothelial barrier breach 220 (e.g., see FIG. 6).Endothelial barrier breach typically results in a first immune responseand vasoconstriction and hemostasis 230 (e.g., see FIG. 7). The secondpathway continues on with a second immune response includingvasodilation, inflammation and proliferation of white blood cells 240(e.g., see FIG. 8). Typically, a third immune response follows the firsttwo, consisting of a repair and remodeling process which producesscarring and fibrosis 250 (e.g., see FIG. 9). This vicious cycle ofchronic insult and injury will continue on, if left unchecked, resultingin chronic degenerative diseases including chronic inflammation andchronic fibrosis 260 (e.g., see FIG. 10). The chronic degenerativedisease results in ischemic cell death including apoptosis 270 (e.g.,see FIG. 11) and necrosis 280 (e.g., see FIG. 12). Along with theseconditions, the ischemic cell injury which results from the sustainedcellular stress caused by the loss of a subject's serum zeta potentialcan lead to metabolic degeneration, epigenetic deregulation, and/orgenetic mutation 290 (e.g., see FIG. 13). In some cases, the breakdownin a subject's cardiovascular system may lead to cancer and metastasis295 (e.g., see FIG. 14). Ultimately, all of this damage can result incatastrophic failure of one or more major organ systems of the subject(e.g., cardiac arrest, stroke, pulmonary embolism) 300 (e.g., see FIG.15). As mentioned loss of serum zeta potential can happen suddenly orslowly over decades. FIG. 16 depicts a pictorial summary of a theory ofchronic degenerative disease over an average human lifespan.

FIGS. 17-18 depict an embodiment of cross-section of a portion of anunhealthy subject's cardiovascular system and a healthy subject'scardiovascular system with a healthy Zeta Shield respectively.

Loss of serum zeta potential leads to Endothelial Erosion (EE), which,when left untreated, eventually results in a variety of conditions asdiscussed herein (e.g., endothelial barrier failure, vasoconstriction,hypertension, hyperlipidemia, Chronic Inflammation, “Auto-Immune”response, scarring, sclerosis, fibrosis, amyloidosis, plaque build-up,etc.). Many of these diseases are associated with aging and accepted asnormal. In some embodiments, many other diseases may be traced back to aloss of serum zeta potential and endothelial erosion. Manynon-communicable diseases (NCDs) associated with aging may be tracedback to a loss of serum zeta potential. NCDs may pose a greater threatto global economic development than fiscal crisis or infectiousdiseases. The lost output caused by the top five NCDs over the next twodecades may exceed forty-seven trillion dollars. FIG. 19 depicts a graphof the average human lifespan for the at least the last several hundredyears and how it has improved and could be improved even more. As can beseen in FIG. 19, once mankind understood how infectious diseases workedand how to prevent and/or treat infectious diseases in the late 19thcentury, the average lifespan roughly doubled. If humanity manages tosolve the problem of non-communicable diseases, the average lifespan ofhumans should increase greatly again. In some embodiments, such NCDs mayinclude: chronic eye diseases (e.g., macular degeneration, retinopathy,cataracts, glaucoma); chronic skin diseases (e.g., systemic sclerosis(scleroderma), psoriasis, dermatitis, eczema, rosacea, lupus); chronicbone, joint, and muscle diseases (e.g., arthritis, osteoporosis,fibromyalgia, degenerative disc disease); chronic gastrointestinaldiseases (e.g., irritable bowel syndrome, inflammatory bowel disease,Chron's disease, leaky gut syndrome, ulcerative colitis); chronic kidneydiseases (e.g., nephritis, nephrosis, nephrotic syndrome, minimal changedisease, focal segmental glomerular sclerosis, membranous nephropathy,end stage renal disease); diabetes and diabetes complications (e.g.,prediabetes, type I and 2 diabetes, peripheral artery disease,peripheral neuropathy, retinopathy, end stage renal disease); chronicrespiratory diseases (e.g., chronic obstructive pulmonary disease;interstitial lung disease, idiopathic pulmonary fibrosis, asthma,emphysema, bronchitis,); cancer (e.g., lung, breast, colorectal,stomach, prostate, kidney, pancreatic, skin, lymphatic, cervical, liver,bladder); cardiovascular diseases (e.g., hypertension, atherosclerosis,heart attack, stroke, peripheral artery disease, erectile dysfunction,chronic fatigue syndrome, metabolic syndrome,); pre-eclampsia andeclampsia; and/or chronic neurological diseases (e.g., dementia,Alzheimer's, Parkinson's, Autism, Epilepsy, multiple sclerosis,amyotrophic lateral sclerosis, peripheral neuropathy, depression,obsessive-compulsive disorder, attention deficit hyperactivity disorder,bipolar disorder). Pharmaceutical compositions and methods describedherein may be used to ameliorate and/or inhibit one or more of thelisted maladies and/or related maladies.

n some embodiments, a method or system described herein may include themeasurement of a carrying capacity of a subject's blood. In someembodiments, a method or system described herein may measure a subject'sdefenses against chronic disease. In some embodiments, a method orsystem described herein may measure a subject's serum zeta potential. Insome embodiments, a method or system described herein may measure alength of a subject's endothelial glycocalyx and/or the amount ofparticular substances known to be shed by a damaged endothelialglycocalyx in the subject's bloodstream. In some embodiments, a methodor system described herein may measure the size of the boundary regionof the endothelial surface layer in a subject's capillaries. Ameasurement of a subject's zeta potential and a subject's endothelialsurface layer health may be used to determine a subject's Zeta Shield.

As regards the referenced hypercoagulation, a method may include ameasurement of the subject's erythrocyte sedimentation rate (ESR). Theerythrocyte sedimentation rate is a blood test which has beentraditionally used to reveal inflammatory activity in the body. When asubject's blood is placed in a tall, thin tube, red blood cells(erythrocytes) gradually clump together and settle to the bottom of thetube. It is thought that inflammation can cause the cells to clump morequickly. Because these clumps are denser than individual cells, theysettle to the bottom more quickly. The sedimentation rate test measuresthe distance red blood cells fall in a test tube in one hour. It isthought that the farther the red blood cells have descended, the greaterthe inflammatory response of a subject's immune system. However, hereinit is known that the increased sedimentation rate may be a result of thereduction or loss of a subject's serum zeta potential and as such thetest might be used to measure a subject's serum zeta potential.

There are several risk factors which may adversely affect a subject'szeta potential. In some embodiments, risk factors may includeelectromagnetic radiation, cigarette smoke, aluminum, heavy metals,radiocontrast agents, sugar, alcohol, poor nutrition, physicalinactivity, bad stress, certain infectious diseases, serious cases oftraumatic physical injury, and aging. In some embodiments, a method orsystem described herein may include administering compositions toinhibit and/or ameliorate adverse effects of one or more risk factors.

As described herein properties of the solution may affect the serum zetapotential. As such the blood serum pH can affect the serum zetapotential. If a subject's pH falls outside of standard parameters acomposition may be administered to a subject to adjust the pH to withinacceptable parameters. The pH of any fluid is the measure of thehydrogen ion (H—) concentration. A pH of 7 is designated as neutral. Thelower the pH, the more acidic the solution. It is accepted that avariety of factors affect blood serum pH, including but not limited towhat is ingested, vomiting, diarrhea, lung function, endocrine function,kidney function, and urinary tract infection. A healthy subject's bloodpH should be between about 7.35 and 7.45. In some embodiments, a methodor system described herein may include administering compositions whichfunction to optimize a subject's serum pH. Compositions designated forthe optimization of blood serum pH may include calcium compounds, silicacompounds, green food powders, silica, sources of bicarbonate,magnesium, chloride, sodium, potassium, olive leaf extract, and/orhorsetail.

In some embodiments, compositions described herein may be formulatedwith a balance of electrolyte compounds designed to be maximallybioavailable, while simultaneously maintaining a high threshold ofsafety for consumption. The ingredients included in compositionsdescribed herein to ensure optimal serum zeta potential includemagnesium chloride, sodium chloride, potassium chloride, olive leafextract, and potassium citrate. The purpose of this composition is toensure the bioavailability of sodium, potassium, chloride, and magnesiumin the GI tract so that these compounds are readily available to ensurethat gains such as increased serum albumin levels and chronic diseasesymptom remission are maintained and not lost.

In some embodiments, a method or system described herein may includecompositions for maintaining and/or increasing a subject's serum zetapotential. The subject's serum zeta potential may be maximized byadministering a composition to a subject which includes one or moresalts. In some embodiments, a composition may include one or morehalogen salts (e.g., chloride salts (e.g., magnesium chloride, potassiumchloride, sodium chloride)). In some embodiments, a composition mayinclude magnesium chloride. In some embodiments, a composition mayinclude magnesium in one or more forms and chloride in one or moreforms. Magnesium chloride may be used because of its bioavailability(i.e., solubility) and low toxicity. Chloride may be administered usingolive plant extracts (e.g., olives, olive oil, olive leaf extract).

In some embodiments, a composition may be administered to a subjectincluding potassium citrate in order to maximize a subject's serum zetapotential. Among other uses, potassium citrate may be used to treatmetabolic problems (e.g., acidosis) caused by kidney disease. In someembodiments, a composition may be administered to a subject includingsodium bicarbonate, potassium bicarbonate, potassium citrate, and/ormagnesium bicarbonate, in order to optimize a subject's serum pH.

In some embodiments, a method or system described herein may includeensuring the health and safety of the endothelial surface layer. In someembodiments, a Zeta Shield of a subject may be maintained and/orrepaired by maintaining and/or repairing a subject's endothelial surfacelayer, including the endothelial glycocalyx. The endothelial glycocalyxmay function to hold the electrokinetic surface charge necessary torepel the plasma proteins and erythrocytes and prevent excessiveperfusion and vascular permeability. The endothelial glycocalyx is thecore component of the endothelial surface layer (ESL), which isgenerated by endothelial cells to modulate contact between abrasiveserum elements and the endothelial cells themselves. The endothelialsurface layer is responsible for vasodilation, which it prompts vianitrous oxide release in response to shear stress, as well as a netnegative electric charge that allows for extremely rapid, low-frictionblood flow and selective endothelial permeability.

Damage to a subject's endothelial surface layer occurs on a regularbasis, due to any sort of wound or infection a subject can suffer. Theendothelial surface layer may be eroded according to the processdescribed herein termed as Endothelial Erosion (EE). A core component ofthe immune response is the enzymatic shedding of parts of theendothelial glycocalyx, which allows for extravasation of lymphocytes tothe site of the wound. In some embodiments, compositions describedherein facilitate healthy interaction between the serum and theendothelial surface layer by supplementing the nutritional intake ofcompounds which make up the endothelial surface layer (including theglycocalyx, which is composed of heparan sulfate, hyaluronic acid,glycosaminoglycans, and proteoglycans).

Compositions may include an endothelial augmentation component (e.g.,Hyaluronic Acid, N-Acetyl Glucosamine, Glucosamine Sulfate, ChondroitinSulfate, or Methylsulfonylmethane) designed to maximize the regenerativecapacity of the endothelial surface layer. Hyaluronic acid is acomponent of the endothelial glycocalyx which may be supplemented incompositions to aid a subject's body's ability to recover from transientconditions in which endothelial surface layer health and function areimpaired or to maximize the endothelial surface layer's capacity toproduce and maintain the negative electric charge which allows it tomaintain selective permeability.

N-Acetyl Glucosarnine (NAG) is a precursor to hyaluronic acid. In someembodiments, compositions may contain NAG or other compounds to augmenta subject's hyaluronic acid production. Glucosamine sulfate is a directprecursor to heparan sulfate, which makes up the side-chains of theprotein strands which comprise the endothelial glycocalyx. Glucosaminesulfate or other compounds may be included in compositions to ensure theavailability of the nutrients needed to heal a damaged endothelialsurface layer. Chondroitin sulfate is a precursor to glycosaminoglycanswhich occur regularly in the endothelial glycocalyx. Chondroitin sulfateor other compounds may be included in described compositions to ensurethe availability of the nutrients needed to heal a damaged endothelialsurface layer.

In some embodiments, the endothelial surface layer may be maintainedand/or repaired by administering a composition including one or moresulfur compounds. Sulfur compounds may include methylsulfonylmethane(MSM), heparan sulfate, glucosarnine sulfate, glucosamine sulfatepotassium, chondroitin sulfate, magnesium sulfate (Epsom salts), and/orN-acetyl glucosamine. The composition(s) may include compounds whichcontribute to the health of and/or form (or contribute to forming) ofthe endothelial glycocalyx including hyaluronic acid and/or collagen.Many of these compounds may be found in what is popularly known as “bonebroth”.

In some embodiments, a method or system described herein may includecompositions for reversing or healing scarring, sclerosis, fibrosis,amyloidosis, calcification and/or plaque build-up which can occur as aresult of the loss of a subject's serum zeta potential and the resultingdamage-referred to as Endothelial Erosion (EE)—to the endothelialsurface layer that occurs as a result of adverse conditions includingbut not limited to the loss of serum zeta potential, physical trauma,infectious disease and/or toxic blood serum contents. Compositions forhealing this damage may include methylsulfonylmethane, amino acids,dextrose, lipids, L-citrulline, DL-Malate, D-Ribose, serrapeptase,nattokinase, tocotrienols (e.g., delta, gamma), N-acetyl cysteine (NAC),vitamins (e.g., B-12, C, D, K), nitric oxide, beet root powder, iron,and/or zinc.

In some embodiments, a method or system described herein may includemaintaining the serum's anti-coagulability and colloid osmotic pressure,resulting in an optimal serum zeta potential. Optimal and/or maximalserum zeta potential will reduce the occurrence of proteinuria.Compositions may include compounds which promote serum maintenance.Compounds may include, for example, methylsulfonylmethane, nattokinase,n-acetyl cysteine, lactoferrin, delta and gamma tocotrienols.Compositions described herein may include a serum maintenance groupcomponent designed to offer protection to the endothelial surface layeragainst damage due to stress, protect the serum itself fromhypercoagulability, and ensure the correct range of serum zetapotential. For example, n-acetyl cysteine is a chemical responsible forvasodilation, which protects the endothelial glycocalyx from harm evenunder such adverse conditions as hyperglycemia. MSM may be included todeliver critical sulfates to the endothelial cells, assisting serum flowby providing a buffer which becomes active at the surface of theendothelial surface layer. Nattokinase is a synthetic fibrinolyticcompound derived from fermented soy, and it helps to ensure thatexcessive coagulability in the serum is discouraged. Lactoferrin is aprotein which may be included in compositions described herein toprovide a reliable, non-glycated source of iron-binding protein whilesimultaneously increasing the concentration of negatively charged serumproteins. Delta & Gamma Tocotrienol (e.g., 90:10) may be included incompositions described herein as a further measure to provideantioxidant support to a subject's body while simultaneously boostingantithrombotic activity.

In some embodiments, a method or system described herein may includecompositions to compensate or replenish bodily resources lost as aresult of a loss of serum zeta potential leading to improper working ofa subject's kidney. Healthy kidneys function to filter out bodily wastesfrom the blood stream but kidneys with a lower serum zeta potential ordamaged endothelial surface layer as discussed herein allow proteins andother essential elements of the body to pass through and out the urinarytract. In some embodiments, a composition may be administered to asubject which includes one or more compounds (e.g., serum albumin,transferrin, lactoferrin, ceruloplasmin, amino acids, dextrose, lipids)typically at depleted levels due to malfunctioning kidneys. In someembodiments, a composition may be administered to a subject whichincludes one or more compounds to be used by a subject's body tonaturally replace elements found at depleted levels due tomalfunctioning kidneys. In some embodiments, a composition may beadministered to a subject which includes one or more compounds (e.g.,Epogen) which prompts a subject's body to naturally replace elementsfound at depleted levels due to malfunctioning kidneys.

In some specific embodiments, a method of treating proteinuria and/orrelated ailments discussed herein may include reestablishing optimallevels of essential plasma proteins lost due to proteinuria. This mayinclude albumin infusions (e.g., 25 g every 8 hours as needed to keepserum albumin levels above 4.0 g/dL). Albumin infusions may make up forurinary losses of albumin, maintain system nutrient transport, and/ormaintain oncotic pressure and limit toxicity, edema and ischemia.

In some specific embodiments, a method of treating proteinuria and/orrelated ailments discussed herein may include reestablishing optimallevels of transferrin lost due to proteinuria. This may includeadministering lactoferrin (e.g., 250 mg every 12 hours as needed to keepserum transferrin levels within an acceptable range) or even moreoptimally administering transferrin itself. Lactoferrin may help asubject's body make up for urinary losses of transferrin (the transportprotein for iron) to limit oxidative stress from unbound iron and irondeficiency anemia.

In some specific embodiments, a method of treating proteinuria and/orrelated ailments discussed herein may include reestablishing maximallevels of a subject's serum zeta potential. In some embodiments, asubject may be administered magnesium chloride (e.g., 1500 mg every 8hours). In some embodiments, a subject may be administeredmethylsulfonylmethane (e.g., 1000 mg every 8 hours). In someembodiments, a subject may be administered potassium citrate (e.g., 100mg every 8 hours). In some embodiments, a subject may be administeredmagnesium chloride, methylsulfonylmethane, and potassium citrate. Insome embodiments, a subject may be administered olive leaf extract(e.g., 750 mg every 12 hours). In some embodiments, a subject may beadministered tocotrienol (e.g., 125 mg, 90% delta, 10% gamma, every 12hours). In some embodiments, a subject may be administered magnesiumchloride, methylsulfonylmethane, olive leaf extract, and tocotrienol.

In some embodiments, a subject's progress may be monitored duringtreatment. The subject's proteinuria may be monitored daily (e.g., byadministering a urine test). As the subject's serum zeta potential isrestored, the level of proteinuria will begin to decrease within daysand will eventually reach the healthy level of albumin elimination(e.g., about 6% of daily albumin production).

In some embodiments, the subject's erythrocyte sedimentation rate may bemonitored. The erythrocyte sedimentation rate will begin to fall withinhealthy parameters as the subject's serum zeta potential is restored.

In some embodiments, a method may include determining a subject's riskfactor quotient by testing for a subject's Zeta Shield. The Zeta Shieldmay consist of a calculation based upon the mathematical combination ofa subject's serum zeta potential score and a second score designed toreflect the health of a subject's endothelial surface layer (e.g., bymeasuring proximal biomarkers including but not limited to a subject'serythrocyte sedimentation rate, glycated albumin vs. total serum albuminand/or hemoglobin hbalc vs. total hemoglobin, blood pressure,circulating glycocalyx fragments, glycocalyx length, the averagedistance of perfusion of erythrocytes into the endothelial surfacelayer, the average endothelial permeability to albumin, etc.).

Dosage and Administration

In some embodiments, chemical compositions described herein may beadministered at a dosage level up to conventional dosage levels, butwill typically be less than about 50mL per day. Suitable dosage levelsfor chemical compositions described herein may be about 0.01 mg to 10 mgper kg body weight of the patient per day, from about 0.1 mg to I mg perkg body weight of the patient per day, or from about 0.01 mg to 0.1 mgper kg body weight of the patient per day. The compound may beadministered on a regimen of up to 6 times per day, between about Ito 4times per day, or once per day.

In the case where an oral composition is employed, a suitable dosagerange is, e.g. from about 0.01 mg to about 10 mg per kg of body weightper day, preferably from about 0.1 mg to about 0.5 mg per kg.

It will be understood that the dosage of the therapeutic agents willvary with the nature and the severity of the condition to be treated,and with the particular therapeutic agents chosen. The dosage will alsovary according to the age, weight, physical condition and response ofthe individual patient. The selection of the appropriate dosage for theindividual patient is within the skills of a clinician.

In addition to administering chemical compositions described herein asdescribed, the compounds may be administered as part of a pharmaceuticalpreparation containing suitable pharmaceutically acceptable carriers,preservatives, excipients and auxiliaries which facilitate processing ofthe chemical compositions described herein which may be usedpharmaceutically. The preparations, particularly those preparationswhich may be administered orally and which may be used for the preferredtype of administration, such as tablets, softgels, lozenges, dragees,and capsules, and also preparations which may be administered rectally,such as suppositories, as well as suitable solutions for administrationby injection or orally or by inhalation of aerosolized preparations, maybe prepared in dose ranges that provide similar bioavailability asdescribed above, together with the excipient. While individual needs mayvary, determination of the optimal ranges of effective amounts of eachcomponent is within the skill of the art.

General guidance in determining effective dose ranges forpharmacologically active compounds and compositions for use in thepresently described embodiments may be found, for example, in thepublications of the International Conference on Harmonisation and inREMINGTON'S PHARMACEUTICAL SCIENCES, 8th Edition Ed. Bertram G. Katzung,chapters 27 and 28, pp. 484-528 (Mack Publishing Company 1990) and yetfurther in BASIC & CLINICAL PHARMACOLOGY, chapters 5 and 66, (LangeMedical Books/McGraw-Hill, New York, 2001).

Pharmaceutical Compositions

Chemical compositions described herein are typically administered orallybut any suitable route of administration may be employed for providing asubject with an effective dosage of drugs of the chemical compositionsdescribed herein. For example, oral, rectal, topical, parenteral,ocular, pulmonary, nasal, and the like may be employed. Dosage formsinclude tablets, troches, dispersions, suspensions, solutions, capsules,creams, ointments, aerosols, and the like. In certain embodiments, itmay be advantageous that the compositions described herein beadministered orally (e.g., tablets, capsules, softgels, solutions,suspensions, etc.).

The compositions may include those compositions suitable for oral,rectal, topical, parenteral (including subcutaneous, intramuscular, andintravenous), ocular (ophthalmic), pulmonary (aerosol inhalation), ornasal administration, although the most suitable route in any given casewill depend on the nature and severity of the conditions being treatedand on the nature of the active ingredient. They may be convenientlypresented in unit dosage form and prepared by any of the methodswell-known in the art of pharmacy.

For administration by inhalation, the drugs used in the presentinvention are conveniently delivered in the form of an aerosol spraypresentation from pressurized packs or nebulizers. The compounds mayalso be delivered as powders which may be formulated, and the powdercomposition may be inhaled with the aid of an insufflation powderinhaler device.

Suitable topical formulations for use in the present embodiments mayinclude transdermal devices, aerosols, creams, ointments, lotions,dusting powders, gels, and the like.

In practical use, drugs used can be combined as the active ingredient inintimate admixtures with a pharmaceutical carrier according toconventional pharmaceutical compounding techniques. The carrier may takea wide variety of forms depending on the form of preparation desired foradministration, e.g., oral or parenteral (including intravenous). Inpreparing the compositions for oral dosage form, pharmaceutical mediamay be employed. For example, in the case of oral liquid preparations(e.g., suspensions, elixirs and solutions) pharmaceutical media mayinclude water, glycols, oils, alcohols, flavoring agents, preservatives,coloring agents and the like. For example, in the case of oral solidpreparations (e.g., powders, capsules and tablets) pharmaceutical mediamay include carriers such as starches, sugars, microcrystallinecellulose, diluents, granulating agents, lubricants, binders,disintegrating agents and the like. In some embodiments, the solid oralpreparations are preferred over the liquid preparations. Because oftheir ease of administration, tablets and capsules represent the mostadvantageous oral dosage unit form in which case solid pharmaceuticalcarriers are obviously employed. If desired, tablets may be coated bystandard aqueous or nonaqueous techniques.

The pharmaceutical preparations may be manufactured in a manner which isitself known to one skilled in the art, for example, by means ofconventional mixing, granulating, dragee-making, softgel encapsulation,dissolving, extracting, or lyophilizing processes. Thus, pharmaceuticalpreparations for oral use may be obtained by combining the activecompounds with solid and semi-solid excipients and suitablepreservatives. Optionally, the resulting mixture may be ground andprocessed. The resulting mixture of granules may be used, after addingsuitable auxiliaries, if desired or necessary, to obtain tablets,softgels, lozenges, capsules, or dragee cores.

The compounds are typically administered in admixture with suitablepharmaceutical diluents, excipients, or carriers (collectively referredto herein as “pharmacologically inert carriers”) suitably selected withrespect to the intended form of administration, that is, oral tablets,capsules, elixirs, syrups and the like, and consistent with conventionalpharmaceutical practices.

Suitable excipients may be fillers such as saccharides (e.g., lactose,sucrose, or mannose), sugar alcohols (e.g., mannitol or sorbitol),cellulose preparations and/or calcium phosphates (e.g., tricalciumphosphate or calcium hydrogen phosphate). In addition binders may beused such as starch paste (e.g., maize or com starch, wheat starch, ricestarch, potato starch, gelatin, tragacanth, methyl cellulose,hydroxypropylmethylcellulose, sodium carboxymethylcellulose, and/orpolyvinyl pyrrolidone). Disintegrating agents may be added (e.g., theabove-mentioned starches) as well as carboxymethyl-starch, cross-linkedpolyvinyl pyrrolidone, agar, or alginic acid or a salt thereof (e.g.,sodium alginate). Auxiliaries are, above all, flow-regulating agents andlubricants (e.g., silica, talc, stearic acid or salts thereof, such asmagnesium stearate or calcium stearate, and/or polyethylene glycol, orPEG). Dragee cores are provided with suitable coatings, which, ifdesired, are resistant to gastric juices. Softgelatin capsules(“softgels”) may be provided with suitable coatings, which, typically,contain gelatin and/or suitable edible dye(s). Animal component-free andkosher gelatin capsules may be particularly suitable for the embodimentsdescribed herein for wide availability of usage and consumption. Forthis purpose, concentrated saccharide solutions may be used, which mayoptionally contain gum arabic, talc, polyvinyl pyrrolidone, polyethyleneglycol (PEG) and/or titanium dioxide, lacquer solutions and suitableorganic solvents or solvent mixtures, including dimethylsulfoxide(DMSO), tetrahydrofuran (THF), acetone, ethanol, or other suitablesolvents and co-solvents. In order to produce coatings resistant togastric juices, solutions of suitable cellulose preparations such asacetylcellulose phthalate or hydroxypropylmethyl-cellulose phthalate,may be used. Dyes or pigments may be added to the tablets or drageecoatings or softgelatin capsules, for example, for identification or inorder to characterize combinations of active compound doses, or todisguise the capsule contents for usage in clinical or other studies.

Other pharmaceutical preparations that may be used orally includepush-fit capsules made of gelatin, as well as soft, thermally sealedcapsules made of gelatin and a plasticizer such as glycerol or sorbitol.The push-fit capsules may contain the active compounds in the form ofgranules that may be mixed with fillers such as, for example, lactose,binders such as starches, and/or lubricants such as talc or magnesiumstearate and, optionally, stabilizers and/or preservatives. In softcapsules, the active compounds may be dissolved or suspended in suitableliquids, such as fatty oils such as rice bran oil or peanut oil or palmoil, or liquid paraffin. In some embodiments, stabilizers andpreservatives may be added.

Liquid dosage forms for oral administration can contain coloring andflavoring to increase subject acceptance. In general, water, a suitableoil, saline, aqueous dextrose (glucose), and related sugar solutions andglycols such as propylene glycol or polyethylene glycols are suitablecarriers for parenteral solutions. Solutions for parenteraladministration preferably contain a water-soluble salt of the activeingredient, suitable stabilizing agents, and if necessary, buffersubstances. Antioxidizing agents such as sodium bisulfite, sodiumsulfite, or ascorbic acid, either alone or combined, are suitablestabilizing agents. Also used are citric acid and its salts and sodiumEDT A. In addition, parenteral solutions can contain preservatives, suchas benzalkonium chloride, methyl- or propyl-paraben, and chlorobutanol.

In some embodiments, an oral composition may include a flavoring. Aflavoring may include something a subject may find palatable. Forexample, a flavoring may include malt extract, Xylitol, Splenda,sucralose or any sweetener. A flavoring may range from 0.01% to 0.10%,0.10% to 1.0%, or 1.0% to 10.0% of a composition.

In some embodiments, a composition may include a colorant. A colorantmay include D&CBlue #1 or any FDA approved color. A colorant may rangefrom 0.001% to 0.010%, 0.010% to 0.10%, or 0.10% to 1.0% of acomposition.

Additional oral compositions which may be used to deliver chemicalcompositions described herein, as well as additional uses, are describedin U.S. Pat. No. 4,666,896 to W amer, Jr. et al., U.S. Pat. No.5,393,516 to Rheinberger et al., and U.S. Pat. No. 5,948,390 to Nelsonet al., as well as U.S. Patent Publication No. 2005/0158252 toRomanowski et al., which are incorporated by reference as if fully setforth herein.

In some embodiments, pulmonary administration of a pharmaceuticalpreparation may be desirable. Pulmonary administration may include, forexample, inhalation of aerosolized or nebulized liquid or solidparticles of the pharmaceutically active component dispersed in andsurrounded by a gas. Possible pharmaceutical preparations, which may beused rectally, include, for example, suppositories, which consist of acombination of the active compounds with a suppository base. Suitablesuppository bases are, for example, natural or synthetic triglycerides,or paraffin hydrocarbons. In addition, it is also possible to usegelatin rectal capsules that consist of a combination of the activecompounds with a base. Possible base materials include, for example,liquid triglycerides, polyethylene glycols, or paraffin hydrocarbons.

Suitable formulations for parenteral administration may include, but arenot limited to, suspensions of the active compounds. Suitable lipophilicsolvents, co-solvents (such as DMSO or ethanol), and/or vehiclesincluding fatty oils, for example, rice bran oil or peanut oil and/orpalm oil, or synthetic fatty acid esters, for example, ethyl oleate ortriglycerides, may be used. Liposomal formulations, in which mixtures ofthe chemical compositions described herein with, for example, egg yolkphosphotidylcholine (E-PC), may be made for injection. Optionally, thesuspension may contain stabilizers, for example, antioxidants such asBHT, and/or preservatives, such as benzyl alcohol.

In some embodiments, an oral composition may include a fragrance.

In some embodiments, a composition may include additional additiveswhich may function in combination or separately from the chemicalcompositions described herein in solution. Additives may function toimprove a subject's health. Additives may include vitamins including,but not limited to, vitamins D and E.

In some embodiments, different compositions may be formulated fordifferent types of users. For professional users (e.g., doctors,veterinarians), compositions may include a greater percentage ofchemical compositions described herein than compositions formulated forover the counter sale to nonprofessionals. Professional compositions maynot include flavorings or colorants.

While previous discussions herein have concentrated on the use ofchemical compositions described herein for treating maladies associatedwith oral cavities of humans and animals. Chemical compositionsdescribed herein may be used for the inhibition and/or amelioration ofvarious maladies associated with humans and/or more particularlyanimals.

While previous discussions herein have concentrated on the use ofchemical compositions and methods described herein for treating maladiesassociated with humans, this example should not be seen as limiting.Compositions described herein may be used to treat other animals (e.g.,mammals) including, but not limited to, felines, canines, avians(birds), reptiles, horses, swine, sheep, goats, deer, tigers, proteinproducing animals (e.g., cattle), and/or lions.

EXAMPLES

Having now described the invention, the same will be more readilyunderstood through reference to the following example(s), which areprovided by way of illustration, and are not intended to be limiting ofthe present invention.

Case Study 1: Zeta Biolongevity, Inc., has developed the initial versionof ZBIO.I which has already been shown to reduce the level ofproteinuria in the ongoing early stages of the initial trial. The firsttest subject, a feline named Claire, had significant proteinuria asshown by her average urine protein/creatinine ratio (UPC), whichwas >0.5 at the beginning of testing, which has diminished significantlyduring the first few months of treatment. As kidney disease is thoughtto be degenerative, the reduction in proteinuria is a promising signthat treatment has been effective.

Claire is a 9-year old female feline who is non-azotemic with severeproteinuria. She has had a chronic upper respiratory infection (URI) formost of her life. The URI, prior to treatment, was very severe-incessantsneezing, heavy mucous drip, congestion, hacking/choking. She was verylethargic and had a poor appetite. Within weeks of treatment, her URIsymptoms have improved significantly. Her breathing is clearer, her nosedoes not run as much, and her appetite has improved substantially.Additionally, Claire's urine PH improved from 7 or higher (outside ofhealthy range) to 6-6.5 (inside healthy range). Claire has significantincrease in energy, sociability and appetite. Claire is entering her 3rdmonth of treatment. It is anticipated she will continue to requireongoing treatment for least 6 more months, due to her long history ofmultiple chronic ailments.

Claire's average proteinuria score before the beginning of trials was0.6667. Since the beginning of treatment, it has declined to 0.2, whichis non-proteinuric according to IRIS guidelines. FIGS. 20-21 depict aspreadsheet showing the preliminary results achieved by treating a catwith severe proteinuria with compositions described herein. The ZetaTheory of Proteinuria & Endothelial Erosion predicts a relationshipbetween serum chemistry and proteinuria.

Claire was administered ZB 10.1 during treatment, the components ofwhich are detailed in TABLE 1 (in some embodiments, percentages in TABLE1 may be +/−10%). ZBIO.I has been developed to ensure that every aspectof Serum Zeta Potential can be maintained. ZB 10.1 ensures that thenegatively charged endothelial surface layer is able to repel abrasiveplasma proteins and erythrocytes throughout the system, but particularlyin the glomerulus. When this relationship breaks down, proteinuria isone result. Hence, a reduction in proteinuria is a significantconfirmation of the underlying theory as well as an indication thatZBIO.I is effective.

TABLE 1 Purpose of Ingredient Maximize mg/day 650 Maximize Health ofMake Up Maximize capsules/day 2 Serum Endothelia for Blood Flow &mg/capsule 325 Zeta 1 Surface Urinary Heal/Repair Ingredient % mg/daymg/capsule Potential Layer Losses Damage Magnesium Chloride 5.0% 32.716.3 X X Potassium Chloride 10.1%  65.3 32.7 X X Sodium Chloride 10.1% 65.3 32.7 X X Potassium Citrate 2.5% 16.3 8.2 X Olive Leaf Extrace15.1%  98.0 49.0 X MSM 15.1%  98.0 49.0 X X Chondroitin sulfate 10.1% 65.3 32.7 X Glucosamine sulfate 10.1%  65.3 32.7 X N-A-G 5.0% 32.7 16.3X Hyaluronic Acid 2.5% 16.3 8.2 X n-Acetyl Cysteine 5.0% 32.7 16.3 X XLactoferrin 5.0% 32.7 16.3 X Tocotrienols 2.5% 16.3 8.2 X Nattokinase2.0% 13.1 6.5 X Totals 100%  650.0 325.0

Case Study 2: In September 2014, Randy Thompson was 54 years old and inexcellent health with none of the lifestyle risk factors or metabolicrisk factors currently presumed to be the root causes of our globalepidemic of chronic disease. Then, on Sep. 19, 2014, Randy Thompsonsuddenly had massive proteinuria. Within two weeks, his serum albumindropped to 1.7 and he was diagnosed with Minimal Change Disease (MCD).Despite high doses of all the usual corticosteroids,anti-inflammatories, immunosuppressants, diuretics, etc., he progressedto End Stage Renal Disease (ESRD) in less than 12 months (Creatinine6.24, BUN 130) and started hemodialysis.

Randy Thompson also developed all the comorbidities associated withdiabetes (peripheral artery disease, peripheral neuropathy, retinopathy,systemic scarring, sclerosis, fibrosis, etc.) without ever havingdiabetes or hyperglycemia. His total cholesterol increased to 550 and hedeveloped over 70 pounds of severe pitting edema.

While on dialysis for 8 months waiting for a kidney transplant (3.5-5.5liters of fluid removed every MWF), Randy Thompson identified theunderlying root cause of proteinuria and the mechanism of progression toESRD and all related metabolic, vascular and neurological comorbidities.To end dialysis Randy Thompson began self-administering magnesiumchloride, methylsulfonylmethane, olive leaf extract, and tocotrienols(delta and gamma). Further Randy Thompson self-administered potassiumcitrate, chondroitin sulfate, glucosamine sulfate, and hyaluronic acid.After dialysis he received albumin infusions to make up for urinarylosses of albumin until his kidneys had a chance to heal. Randy Thompsonbegan self-administering lactoferrin to ameliorate severe irondeficiency anemia due to urinary losses of transferrin.

After months of treatment, it became apparent that Randy Thompson hadbeen able to reverse and cure his ESRD and all the associatedcomorbidities. Randy Thompson has now fully recovered and has normalkidney function once again. Randy Thompson has also restored all theblood flow to his legs, ankles and feet and has healed all theperipheral nerve damage.

In this patent, certain U.S. patents, U.S. patent applications, andother materials (e.g., articles) have been incorporated by reference.The text of such U.S. patents, U.S. patent applications, and othermaterials is, however, only incorporated by reference to the extent thatno conflict exists between such text and the other statements anddrawings set forth herein. In the event of such conflict, then any suchconflicting text in such incorporated by reference U.S. patents, U.S.patent applications, and other materials is specifically notincorporated by reference in this patent.

Further modifications and alternative embodiments of various aspects ofthe invention will be apparent to those skilled in the art in view ofthis description. Accordingly, this description is to be construed asillustrative only and is for the purpose of teaching those skilled inthe art the general manner of carrying out the invention. It is to beunderstood that the forms of the invention shown and described hereinare to be taken as the presently preferred embodiments. Elements andmaterials may be substituted for those illustrated and described herein,parts and processes may be reversed, and certain features of theinvention may be utilized independently, all as would be apparent to oneskilled in the art after having the benefit of this description of theinvention. Changes may be made in the elements described herein withoutdeparting from the spirit and scope of the invention as described in thefollowing claims.

What is claimed:
 1. A therapeutic composition to alleviate proteinuriain a human patient by reducing permeability of the vascular endothelialsurface layer comprising: 1) an oral dose of a component of vascularendothelial glycocalyx comprising hyaluronic acid; 2) an oral dose oflactoferrin in a therapeutically effective amount to increase serumlevels of transferrin; 3) an oral dose of a pharmaceutically effectiveamount of chondroitin and glucosamine, wherein the effective amountincreases the serum zeta potential by reducing hypercoagulation oferythrocytes; and 4) a sulfur compound selected from the groupconsisting of methylsulfonylmethane (MSM), heparan sulfate, magnesiumsulfate, N-acetyl glucosamine, a sulfur derivative of the chondroitin, asulfur derivative of the glucosamine, and combinations thereof.
 2. Thecomposition of claim 1, wherein the sulfur compound ismethylsulfonylmethane to ameliorate damage to the endothelial surfacelayer.
 3. The composition of claim 1 further tocotrienols.
 4. Thecomposition of claim 3, wherein the tocotrienols are comprised of deltaand gamma tocotrienols in a 90:10 ratio respectively.
 5. The compositionof claim 1, further comprising a compound selected form the groupconsisting of nattokinase, potassium citrate, and olive leaf extract andcombinations thereof.
 6. The composition of claim 1 further comprisingN-acetyl cysteine.
 7. The composition of claim 1 further comprisingmagnesium.
 8. The composition of claim 1, wherein the hyaluronic acid,glucosamine, chondroitin and lactoferrin are combined in an oral formhaving a predetermined unit dose effective to decrease excess urinaryprotein excretion.
 9. The composition of claim 1, further comprising anintravenous dose of albumin.